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dc.contributor.authorHoughton, RLen
dc.contributor.authorReed, DEen
dc.contributor.authorHubbard, MAen
dc.contributor.authorDillon, MJen
dc.contributor.authorChen, Hen
dc.contributor.authorCurrie, BJen
dc.contributor.authorMayo, Men
dc.contributor.authorSarovich, DSen
dc.contributor.authorTheobald, Ven
dc.contributor.authorLimmathurotsakul, Den
dc.contributor.authorWongsuvan, Gen
dc.contributor.authorChantratita, Nen
dc.contributor.authorPeacock, SJen
dc.contributor.authorHoffmaster, ARen
dc.contributor.authorDuval, Ben
dc.contributor.authorBrett, PJen
dc.contributor.authorBurtnick, MNen
dc.contributor.authorAucoin, DPen
dc.date.accessioned2021-09-16T14:36:01Z
dc.date.available2021-09-16T14:36:01Z
dc.date.issued2014-03-20en
dc.identifier.issn1935-2727en
dc.identifier.urihttp://hdl.handle.net/10026.1/17807
dc.description.abstract

Burkholderia pseudomallei is a soil-dwelling bacterium and the causative agent of melioidosis. Isolation of B. pseudomallei from clinical samples is the "gold standard" for the diagnosis of melioidosis; results can take 3-7 days to produce. Alternatively, antibody-based tests have low specificity due to a high percentage of seropositive individuals in endemic areas. There is a clear need to develop a rapid point-of-care antigen detection assay for the diagnosis of melioidosis. Previously, we employed In vivo Microbial Antigen Discovery (InMAD) to identify potential B. pseudomallei diagnostic biomarkers. The B. pseudomallei capsular polysaccharide (CPS) and numerous protein antigens were identified as potential candidates. Here, we describe the development of a diagnostic immunoassay based on the detection of CPS. Following production of a CPS-specific monoclonal antibody (mAb), an antigen-capture immunoassay was developed to determine the concentration of CPS within a panel of melioidosis patient serum and urine samples. The same mAb was used to produce a prototype Active Melioidosis Detect Lateral Flow Immunoassay (AMD LFI); the limit of detection of the LFI for CPS is comparable to the antigen-capture immunoassay (∼0.2 ng/ml). The analytical reactivity (inclusivity) of the AMD LFI was 98.7% (76/77) when tested against a large panel of B. pseudomallei isolates. Analytical specificity (cross-reactivity) testing determined that 97.2% of B. pseudomallei near neighbor species (35/36) were not reactive. The non-reactive B. pseudomallei strain and the reactive near neighbor strain can be explained through genetic sequence analysis. Importantly, we show the AMD LFI is capable of detecting CPS in a variety of patient samples. The LFI is currently being evaluated in Thailand and Australia; the focus is to optimize and validate testing procedures on melioidosis patient samples prior to initiation of a large, multisite pre-clinical evaluation.

en
dc.format.extente2727 - ?en
dc.format.mediumElectronic-eCollectionen
dc.languageengen
dc.language.isoengen
dc.subjectHumansen
dc.subjectBurkholderia pseudomalleien
dc.subjectMelioidosisen
dc.subjectPolysaccharides, Bacterialen
dc.subjectAntibodies, Bacterialen
dc.subjectAntibodies, Monoclonalen
dc.subjectAntigens, Bacterialen
dc.subjectChromatography, Affinityen
dc.subjectSensitivity and Specificityen
dc.subjectPoint-of-Care Systemsen
dc.subjectThailanden
dc.subjectAustraliaen
dc.titleDevelopment of a prototype lateral flow immunoassay (LFI) for the rapid diagnosis of melioidosis.en
dc.typeJournal Article
plymouth.issue3en
plymouth.volume8en
plymouth.publication-statusPublisheden
plymouth.journalPLoS neglected tropical diseasesen
dc.identifier.doi10.1371/journal.pntd.0002727en
plymouth.organisational-group/Plymouth
plymouth.organisational-group/Plymouth/Faculty of Health
plymouth.organisational-group/Plymouth/Faculty of Health/Peninsula Medical School
plymouth.organisational-group/Plymouth/Users by role
plymouth.organisational-group/Plymouth/Users by role/Academics
dc.identifier.eissn1935-2735en
dc.rights.embargoperiodNot knownen
rioxxterms.versionofrecord10.1371/journal.pntd.0002727en
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserveden
rioxxterms.typeJournal Article/Reviewen


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